江俊斌CHIANG, CHUN-PIN臺灣大學:臨床牙醫學研究所林佳蓉Lin, Chia-JungChia-JungLin2010-05-262018-07-092010-05-262018-07-092009U0001-1606200902200000http://ntur.lib.ntu.edu.tw//handle/246246/184175背景和目的：在上皮癌入侵的過程中，穩定的上皮細胞會出現類似間葉細胞的形態轉變，此過程稱為EMT(epithelial-mesenchymal transition)。具有淋巴組織的特異性的標記– podoplanin – 牽涉到許多人類上皮癌的腫瘤入侵，因此在本實驗中，我們研究了podoplanin與 “鈣黏著素E“(E-cadherin)在正常黏膜、口腔上皮變異、及口腔鱗狀細胞癌的表現來了解他們與腫瘤入侵、臨床病理特徵、及病人存活狀況的相關性。料和方法：透過免疫組織化學染色方法來評估podoplanin與鈣黏著素E於正常口腔黏膜、口腔上皮變異、口腔鱗狀細胞癌的表現。計算podoplanin及鈣黏著素E在正常口腔黏膜、口腔上皮變異、口腔鱗狀細胞癌細胞、及口腔鱗狀細胞癌腫瘤內外的淋巴管密度的標記分數，並評估此分數與臨床病理特徵及口腔鱗狀細胞癌病人存活的相關性。果： 我們發現腫瘤周圍的的淋巴管密度的減少與較大的腫瘤(P = 0.000)、遠端轉移(P = 0.023)、及較高的臨床分級(P = 0.001)有顯著關係。除此之外，腫瘤內的平均淋巴管密度的減少也與較大的腫瘤(P = 0.010) 顯著相關。在口腔鱗狀細胞癌的病人中，腫瘤周圍淋巴管密度小於34條血管/每100倍視野者，比大於等於34條血管/每100倍視野者有明顯較差的全存活率(P = 0.036)及疾病特異存活率(P = 0.020)。同時，在口腔鱗狀細胞癌病人中podoplanin的表現有顯著增加，並與局部淋巴轉移(P = 0.026)、遠端轉移(P = 0.021)、及較差的組織分化(P = 0.013)相關。且在口腔鱗狀細胞癌的病人中，podoplanin標計分數大於等於20%者，比小於20%者有明顯較差的全存活率(P = 0.033)。鈣黏著素E的減少在口腔鱗狀細胞癌病人與較年輕的族群(P = 0.002)、局部淋巴轉移(P = 0.016)、及較差的組織分化(P = 0.001)顯著相關。在口腔鱗狀細胞癌的病人中，podoplanin標計分數小於150%者，比大於等於150%者有明顯較差的全存活率(P = 0.016)及疾病特異存活率(P = 0.005)。而且在podoplanin表現高的腫瘤侵入前緣(invasion front)則發現鈣黏著素E明顯減少。論：腫瘤周圍的淋巴管密度可用做生物標記來預測口腔鱗狀細胞癌病人的腫瘤大小、臨床分級和預後。在口腔癌化的過程中發現有podoplanin表現的明顯增加及鈣黏著素E的減少，且與口腔鱗狀細胞癌病人的淋巴轉移及較差的預後相關。這顯示podoplanin及鈣黏著素E的標記分數也許可作為預測口腔鱗狀細胞癌的淋巴轉移及預後的有效生物標記。在大部分的口腔鱗狀細胞癌中，podoplanin鈣黏著素E可能是以EMT的方式參與腫瘤入侵，然而口腔鱗狀細胞癌入侵機制仍需要更進一步的研究。Background and purposes: The transformation from stable epithelial cell to the mesenchymal phenotype, named as epithelial-mesenchymal transition (EMT), was noted in the process of carcinoma invasion. The podoplanin, a specific lymphatic marker, is involved in the process of the cancer invasion in a variety of human carcinomas. In this study, we investigated the expression of podoplanin and E-cadherin in the normal oral mucosa (NOM), oral epithelial dysplasia (OED), and oral squamous cell carcinoma (OSCC) samples to understand their association with tumor invasion, clinicopathological parameters, and patients’ survival. aterials and methods: The expression of podoplanin and E-cadherin in NOM, OED, and OSCC samples was evaluated by immunohistochemistry. The podoplanin and E-cadherin labeling scores (LSs) for NOM, OED, and OSCC samples and the intratumoral and peritumoral lymphatic vessel densities (iLVD and pLVD) in OSCC samples were counted and correlated with clinicopathological parameters and survival of OSCC patients. esults: We found that there was a significant reduction of the mean pLVD in OSCCs with larger tumor size (P = 0.000), with distant metastasis (P = 0.023), and with more advanced clinical stage (P = 0.001). In addition, there was also a significant decrease of the mean iLVD in OSCCs with larger tumor size (P = 0.010). OSCC patient with the pLVD < 34 vessels/100× field had a significantly poorer overall (P = 0.036) and disease-specific survival (P = 0.020) than those with the pLVD > 34 vessels/100× field. A significant increase in podoplanin expression was found in OSCCs with regional lymph node metastasis (P = 0.026), distant metastasis (P = 0.021), and less histologic differentiation (P = 0.013). Moreover, OSCC patient with the podoplanin LS > 20% had a significantly poorer overall survival than those with the podoplanin LS < 20% (P = 0.033). There was a significant decrease in E-cadherin expression in younger OSCC patients (P = 0.002) and in OSCCs with regional lymph node metastasis (P = 0.016) and less histologic differentiation (P = 0.001). OSCC patient with the E-cadherin LS < 150% had a significantly poorer overall (P = 0.016) and disease-specific survival (P = 0.005) than those with the E-cadherin LS > 150%. Furthermore, cancer cells in the invasion front with high podoplanin expression showed a significant loss of E-cadherin.onclusions: The pLVD may act as a biomarker to predict the T status, clinical stage, and prognosis of OSCC patients. An increase of podoplanin expression and a loss of E-cadherin expression are noted during oral carcinogenesis and are associated with more lymph node metastasis and poorer prognosis of OSCC patients. This suggests that both podoplanin and E-cadherin LSs may be useful biomarkers for prediction of tumor metastasis in OSCCs and prognosis of OSCC patients. The podoplanin and E-cadherin may be involved in cancer invasion with EMT in most OSCCs. However, further studies on the mechanisms of OSCC invasion are needed.國立臺灣大學碩士學位論文 口試委員會審定書 .................................................1誌 ...............................................................................................................6文摘要.........................................................................................................7bstract ........................................................................................................9ntroduction ...............................................................................................11iterature Review ........................................................................................13. Tumor invasion .................................................................................13I. Tumor lymphatic system ...................................................................15II. E-cadherin ........................................................................................19V. Podoplanin ........................................................................................21urposes of this study .................................................................................28aterials and Methods ..........................................................................29. Patients and specimens ...................................................................29I. Immunostain ........................................................................................31II. Scoring ...............................................................................................33V. Statistical analysis ..........................................................................34esults ......................................................................................................36. Correlations between lymphatic vessel density in OSCC samples and clinicopathological parameters of OSCC patients ........................................36I. Podoplanin and E-cadherin expressions in NOM, OED, and OSCC samples ....................................................................................................................37II. Correlations between podoplanin or E-cadherin expression in OED samples and clinicopathological parameters of OED patients ...................38V. Correlations between podoplanin or E-cadherin expression in OSCC samples and clinicopathological parameters of OSCC patients ...................38. Correlation between podoplanin and E-cadherin expression ............39iscussion ...............................................................................................40. Correlations between lymphatic vessel density and clinicopathological parameters of OSCC patients ...................................................................40I. Podoplanin expression in NOM, OED, and OSCC samples ...................41II. E-cadherin expression in NOM, OED, and OSCC samples ...................44V. Correlation between podoplanin and E-cadherin expression in OSCC samples ......................................................................................................45onclusions ...............................................................................................47eferences ...............................................................................................48ables ......................................................................................................67igure legends ........................................................................................74igures ......................................................................................................78application/pdf8271899 bytesapplication/pdfen-USpodplanin鈣黏著素E腫瘤入侵淋巴管密度口腔鱗狀細胞癌podoplaninE-cadherintumor invasionlymphatic vessel densityoral squamous cell carcinoma[SDGs]SDG3http://ntur.lib.ntu.edu.tw/bitstream/246246/184175/1/ntu-98-R96422002-1.pdf